title: Identification of the human testis protein phosphatase 1 interactome.
abstract: Protein phosphorylation is a critical regulatory mechanism in cellular signalling. To this end, PP1 is a major eukaryotic serine/threonine-specific phosphatase whose cellular functions, in turn, depend on complexes it forms with PP1 interacting proteins-PIPs. The importance of the testis/sperm-enriched variant, PP1γ2, in sperm motility and spermatogenesis has previously been shown. Given the key role of PIPs, it is imperative to identify the physiologically relevant PIPs in testis and sperm. Hence, we performed Yeast Two-Hybrid screens of a human testis cDNA library using as baits the different PP1 isoforms and also a proteomic approach aimed at identifying PP1γ2 binding proteins. To the best of our knowledge this is the largest data set of the human testis PP1 interactome. We report the identification of 77 proteins in human testis and 7 proteins in human sperm that bind PP1. The data obtained increased the known PP1 interactome by reporting 72 novel interactions. Confirmation of the interaction of PP1 with 5 different proteins was also further validated by co-immunoprecipitation or protein overlays. The data here presented provides important insights towards the function of these proteins and opens new possibilities for future research. In fact, such diversity in PP1 regulators makes them excellent targets for pharmacological intervention.
authors.affiliation: Signal Transduction Laboratory, Centre for Cell Biology, Biology Department, University of Aveiro, 3810-193 Aveiro, Portugal. mfardilha@ua.pt
authors.forename: Margarida
authors.initials: M
authors.lastname: Fardilha
authors.forename: Sara L C
authors.initials: SL
authors.lastname: Esteves
authors.forename: Luís
authors.initials: L
authors.lastname: Korrodi-Gregório
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Vintém
authors.forename: Sara C
authors.initials: SC
authors.lastname: Domingues
authors.forename: Sandra
authors.initials: S
authors.lastname: Rebelo
authors.forename: Nick
authors.initials: N
authors.lastname: Morrice
authors.forename: Patricia T W
authors.initials: PT
authors.lastname: Cohen
authors.forename: Odete A B
authors.initials: OA
authors.lastname: da Cruz e Silva
authors.forename: Edgar F
authors.initials: EF
authors.lastname: da Cruz e Silva
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D018076:DNA, Complementary; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D015723:Gene Library; D006801:Humans; D008297:Male; D011485:Protein Binding; D020033:Protein Isoforms; D054645:Protein Phosphatase 1 / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013081:Sperm Motility / Q000502:physiology; D013737:Testis / Q000201:enzymology*; D020798:Two-Hybrid System Techniques
chemical_list: D018076:DNA, Complementary; D020033:Protein Isoforms; D054645:Protein Phosphatase 1